Chromatographic purification of Biomolecules and their characterization - CHROBIO

Aim of the proposed research is the design of non-invasive chromatographic methods suitable for purification and concentration of biomolecules (viruses and proteins) and their characterization. Purification and analysis methods will be focused on measles virus. Measles virus is used as a vaccine and is found very suitable as a gene vector and as an oncolytic virotherapeutic, with the last two applications requiring large amounts of the virus. All biotechnological products are subjected to rigorous quality requirements to ensure their safe usage.

Therefore, non-invasive purification methods upgrading their quality and sustainable production and ensuring stability of these fragile biomolecules are essential. This project will base purification on chromatography using monolith stationary phases which are featured by their suitability for biomolecules due to high porosity, high binding capacity and convective based transport. Ion-exchange and affinity chromatography will be employed and various conditions tested for obtaining target material with high purity and yield. Also, various formulation conditions enabling preservation of virus viability and protein functionality and stability throughout the process will be tested.

The analysis of the virus infectivity and protein functionality, removal of contaminants and the overall procedure effectiveness will be done by classical biochemical and virological methods as well as by some novel methods such as nanoparticle analysis that enables detection of virus and protein aggregates (NanoSight).

Application of the research

Results of the project are expected to provide data about measles contributing to fundamental knowledge about the virus biochemistry but also enabling upgrade of the virus production in biotechnology.

Project category

HRZZ Installation Research Projects

Project budget

679.000,00 HRK

Starting date

15.09.2014.

Finishing date

14.09.2017.